The principal overall objective of the proposed research is the development of gas phase techniques, gas chromatography and combined gas chromatography-mass spectrometry-computer systems, for the analysis of intact glucuronides in biologic fluids and tissues. Human urine, obtained from persons of varying age, sex, and genetic background, will be used initially as the source of glucuronides for this study. Quantitative procedures for extraction or isolation of glucuronide mixtures from urine will be established. An attempt will be made to utilize crude urine for the analysis of glucuronides. The gas phase properties of various derivatives, such as permethyl, silyl, and acetyl, will be studied; those derivatives having the most desirable properties will be used in the routine analyses. A comprehensive study of the parameters needed for high resolution gas chromatographic separation of glucuronide mixtures will be carried out. Mass spectra of the various derivatives of urinary glucuronides will be used for identification of these compounds. Single and multiple ion monitoring techniques will be developed to specifically detect glucuronides in complex mixtures. Quantification will be carried out by peak area comparison to suitable standards or by multiple ion monitoring with labelled analogs as reference compounds. The techniques which will be developed will be applied to the separation and identification of other compounds in urine, especially those which reflect enzyme defects and can cause mental or physical retardation.